Introduction: Cancer is describes as the disease that results due to cellular changes and these changes cause the uncontrolled growth and division of cells. A cell receives instructions to die so that the body can replace it with a newer cell that functions better. Cancerous cells lack the components that instruct them to stop dividing and to die. Chemotherapeutic drugs (CDs) are the most widespread worldwide modality used in cancer treatment, and other autoimmune diseases. However, their non-selective mechanism of action affects both cancerous and non-cancerous cells, that resulting in well documented side effects. Nurses are at risk of suffering side effects. Little negligence or mistake may lead to adverse unpleasant effects for patients, staff and environment. This study was undertaken to assess the knowledge and attitude regarding safe handling of chemotherapeutic drugs among nursing personnel in tertiary care hospital, India. Aim: The aim is to assess the knowledge and attitude regarding safe handling of chemotherapeutic drugs among nursing personnel. Material & Methods: Non-experimental descriptive cross sectional survey approach was used. Through convenient sampling, 60 subjects were selected. The knowledge questionnaire to assess knowledge and Likert’s scale to assess attitude was used. Descriptive statistics, pearson corelation and ANOVA /t- tests were used to analysis data. Results: The result revealed that the majority of nursing personnel (55%) had poor knowledge and (46.7%) had mild positive attitude regarding safe handling of chemotherapeutic drugs. The statistically significant association was found between knowledge regarding safe handling of chemotherapeutic drugs and gender, education of nursing personnel. The statistically highly significant association was found between attitude regarding safe handling of chemotherapeutic drugs and oncology work experience of nursing personnel. Conclusion: The results of the present study reveal that there is poor knowledge and mild positive attitude regarding safe handling of chemotherapeutic drugs among nursing personnel of tertiary care hospital, India.